Institute: Oklahoma
Year Established: 2003 Start Date: 2003-03-01 End Date: 2005-02-28
Total Federal Funds: $50,000 Total Non-Federal Funds: $100,000
Principal Investigators: Gilbert John, Mario Rivera, Gary Yen
Project Summary: Since September 11th, Homeland Security in the United States has become more important, as many aspects of security in this country are being examined and developed. One aspect is the security of drinking water. Deliberate contamination of drinking water make it imperative to have an efficient, sensitive, specific and rapid sensor that can detect both xenobiotics and microbial organisms that can cause harm to individuals. Billions of dollars are being made available from government and state agencies to develop systems that can continuously monitor drinking water. A multi-discipline group at Oklahoma State University is involved in developing a dual sensor that can be used in this capacity. Our proposal specifically addresses two critical areas that are important for further development of a dual sensor that can detect potentially harmful xenobiotics (toxicants) and pathogenic bacteria in water. The first area specifically addresses the issue of having stable proteins that can be maintain their function under various environmental conditions. The cytochrome P450 protein from the human liver is normally involved in detoxifying and toxifying a broad range of xenobiotics, thereby the protein can be used to directly link xenobiotics to human toxicity. A number of isoforms are present in the liver, but some of these proteins are not stable (CYP3A4), compared to stable proteins (CYP1A2). Therefore, our proposal seeks to develop a method of improving stability of CYP 3A4 using molecular modeling, molecular biology, and spectrophotometric techniques that will increase ion-pair interactions in the protein. The second area addresses the specificity of autofluorescence signatures (spectrofluorimetry) from bacteria, as a large collection of different types of bacteria will be tested. Having methods than can improve the stability of cytochrome P450 without compromising function is critical for generating unique spectra that can be used to specifically identify potentially harmful toxicants. In addition, demonstrating that bacteria have unique autofluorescence signatures can provide a powerful method for identifying different types of bacteria. The conclusion of this study will enable our group to submit a more extensive grant that will develop a working dual sensor that can be used to detect both xenobiotics and pathogenic bacterial in drinking water.